Mouse embryos at E15 were used for in vivo labeling of annexin V Alexa 488 conjugation (A13201; Thermo Fisher Scientific). Brain or other organs (heart, intestine) were directly incubated with 20 μl annexin V in 400 μl annexin V reaction buffer (25 mM Hepes, 140 mM NaCl, and 1 mM EDTA, pH 7.4) in 1.5-ml tubes for 15 min at room temperature. After three times wash by PBS, samples were fixed by 4% paraformaldehyde in 0.1 M phosphate buffer for 1 h at room temperature. Samples were further incubated with 20% and 30% sucrose in PBS for 12 h. 10 μm cryosections were prepared by Leica CM1850 (Leica). Sections were stained with DAPI (D523; DOJINDO) and coverslipped. Images were obtained by confocal microscopy (FV1200IXGP44; Olympus).
A13201
Manufactured by Thermo Fisher Scientific
Sourced in United States
The A13201 is a laboratory centrifuge that can be used for sample separation and preparation. It has a rotor capacity of up to 6 tubes and can achieve a maximum speed of 6,000 rpm. The centrifuge is designed for basic benchtop use in research and clinical laboratories.
Automatically generated - may contain errors
Lab products found in correlation
Cm1850, by Leica (1 mentions)
Dapi d523, by Dojindo Laboratories (1 mentions)
Facscanto 2, by BD (1 mentions)
Annexin 5 alexa fluor 488, by Thermo Fisher Scientific (1 mentions)
Fxcycle pi rnase staining solution, by Thermo Fisher Scientific (1 mentions)
Annexin 5 fitc, by Thermo Fisher Scientific (1 mentions)
Hoechst 33342, by Thermo Fisher Scientific (1 mentions)
Dmi4000b, by Leica (1 mentions)
P3566, by Thermo Fisher Scientific (1 mentions)
Cytoflex flow cytometry, by Beckman Coulter (1 mentions)
Cytexpert software, by Beckman Coulter (1 mentions)
6 protocols using a13201
1
In vivo Annexin V Labeling of Mouse Embryos
2
Apoptosis and Cell Cycle Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were incubated with NAC and VitC for 48 h and washed in PBS. For apoptosis analysis, 5 mL Alexa Fluor 488 Annexin V (A13201, ThermoFisher) and 1 mL propidium iodide (PI) working solution (100 mg/mL, 421301, Biolegend) was added to 100 mL cell suspension. After incubating 15 min, 400 mL 1X Annexin-binding buffer was added and the cells were analyzed by flow cytometry (BD FACSCanto II; excitation wavelength 488 nm, and emission 530/30 nm). For cell cycle analysis, fixed cells were incubated with FxCycle PI/RNase Staining Solution (1985233, Invitrogen) for 30 min at room temperature, and then analyzed by flow cytometry (excitation wavelength 488 nm, and emission 530/30 nm).
3
Apoptosis Evaluation by Flow Cytometry
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Apoptotic cells were evaluated by ANXA5 and propidium iodide (PI) staining (Invitrogen, A13201) according to the manufacturer’s instructions, and analyzed by flow cytometry (Beckman Coulter, USA).
4
Quantifying Apoptosis via Flow Cytometry
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Apoptosis was assayed via Annexin V-FITC and propidium iodide (PI) staining according to the manufacturer's protocol (Invitrogen Carlsbad, CA, USA, Catalogue number, A13201). HCT116 cells were treated with CX-5461 or vehicle for 72 h; then the percentage of apoptotic cells was analysed by flow cytometry. Early apoptotic cells are Annexin V positive and PI negative (lower right quadrant in FACS profile); late apoptotic and dead cells are Annexin V and PI positive (upper right quadrant). 10,000 cells were counted for each sample.
5
Apoptosis Quantification via Annexin V
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
At 2 h postinjury, cells were stained with a mixture that included annexin V conjugated to Alexa-Fluor 488 (1:50 dilution from stock; A13201, Invitrogen), and 5 μg/mL Hoechst 33342 (Invitrogen). The stain mixture, in annexin binding buffer, was added directly to the 250 μL media in each well. Cells were incubated in the dark for 15 min and 5 images per well were taken on a Leica DMI4000B (Wetzlar, Germany) fluorescence microscope at 10× magnification. ImageJ software (National Institutes of Health, Bethesda, MD) was used to count Hoechst-positive and annexin-positive cells. Hoechst was used to determine the total number of cells, and percentage of annexin V–positive cells was determined from approximately 600 total cells per well.
6
Apoptosis Quantification via Flow Cytometry
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
After 24 h of treatment, apoptosis was evaluated by double labeling cells with Annexin V/Alexa Fluor 488 conjugate (A13201, Invitrogen, USA) and propidium iodide (PI) (P3566, Invitrogen, USA). During early apoptosis, Annexin V stains phosphatidylserine (PS) in the outer membrane. PI can enter the cell and attach to double-stranded DNA when the cell membrane becomes permeable. Cells in the late apoptosis stage were stained with both Annexin V and PI. The apoptotic cells were quantified with CytoFLEX flow cytometry and CytExpert software (Version 2.4.0.28) (Beckman Coulter, USA) and calculated as percentages using GraphPad Prism9 software.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!